LOCUS       Exported                8330 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pMK500-TEV, complete sequence.
ACCESSION   AY647986
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8330)
  AUTHORS   Guo D, Hazbun TR, Xu XJ, Ng SL, Fields S, Kuo MH.
  TITLE     A tethered catalysis, two-hybrid system to identify protein-protein 
            interactions requiring post-translational modifications
  JOURNAL   Nat. Biotechnol. 22 (7), 888-892 (2004)
  PUBMED    15208639
REFERENCE   2  (bases 1 to 8330)
  AUTHORS   Kuo M-H., Xu X-J., Ng S-L.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2004) Biochem. Mol. Biol., Michigan State 
            University, 309 BCH Building, East Lansing, MI 48824, USA
REFERENCE   3  (bases 1 to 8330)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8330
                     /organism="Yeast two-hybrid vector pMK500-TEV"
                     /mol_type="genomic DNA"
                     /label=tethered catalysis bait construct
                     /note="tethered catalysis bait construct"
                     /db_xref="taxon:284157"
     CDS             434..2104
                     /codon_start=1
                     /product="GDBD-TEV-CTDx3-Kin28-HA fusion protein"
                     /label=GDBD-TEV-CTDx3-Kin28-HA fusion protein
                     /note="derived from Saccharomyces cerevisiae; S5 of the CTD
                     is phosphorylated by the tethered Kin28"
                     /protein_id="AAT97348.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSNYLFDDEDTPPNPKKE
                     IELENLYFQGEFQYSPTSPSYSPTSPSYSPTSPSLTTMAMKVNMEYTKEKKVGEGTYAV
                     VYLGCQHSTGRKIAIKEIKTSEFKDGLDMSAIREVKYLQEMQHPNVIELIDIFMAYDNL
                     NLVLEFLPTDLEVVIKDKSILFTPADIKAWMLMTLRGVYHCHRNFILHRDLKPNNLLFS
                     PDGQIKVADFGLARAIPAPHEILTSNVVTRWYRAPELLFGAKHYTSAIDIWSVGVIFAE
                     LMLRIPYLPGQNDVDQMEVTFRALGTPTDRDWPEVSSFMTYNKLQIYPPPSRDELRKRF
                     IAASEYALDFMCGMLTMNPQKRWTAVQCLESDYFKELPPPSDPSSIKIRNVMAIPGGGR
                     IFYPYDVPDYAGYPYDVPDYAGSYPYDVPDYAAQCGRSS"
     CDS             434..874
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             932..952
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    962..1024
                     /note="trimeric carboxyl terminal domain repeat of RNA 
                     polymerase II"
     misc_feature    1040..1957
                     /label=Kin28
                     /note="Kin28"
     CDS             1991..2080
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     terminator      2519..2706
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    3538..4700
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     rep_origin      3863..4700
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1/ARS416"
     CDS             complement(4539..5213)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(5214..5315)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        5421..5525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5526..6386
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6557..7145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"